Drosophila Melanogaster BAC Library
The CHORI-221 Drosophila
melanogaster BAC library has been constructed in our laboratory
by Kazutoyo Osoegawa
in collaboration with the Berkeley
Drosophila Genome Project headed by Gerald M. Rubin and
co-directed by Susan E. Celniker at Lawrence Berkeley National
Laboratory. High-molecular-weight DNA from an isogenic y1;
cn1 bw1 sp1 strain (B.J. Brizuela et al., 1994, Genetics 137:
803; Bloomington stock number 2057) was prepared by Roger
A. Hoskins. The agarose embedded DNA was sheared by repeating
freezing and thawing. The fragmented ends were polished by
subsequent treatments with Mung bean nuclease and T4 DNA polymerase,
respectively. The polished ends were ligated to the blunt-end
side of an adapter which has a 3'overhang. Size fractionated
DNA was cloned into the pTARBAC6 vector between the BstXI
sites. The ligation products were transformed into DH10B electrocompetent
cells (BRL Life Technologies). The library has been arrayed
into 384-well microtiter dishes and also gridded onto a 22x22cm
nylon high-density filter for screening by probe hybridization.
The hybridization membrane represents over 18,000 distinct
D. melanogaster BAC clones, stamped in duplicate.
data for CHORI-221 Drosophila melanogaster BAC Library:
120 Mb euchromatic portion)
The total library
should represent approximately 15-fold total genomic representation.
clone average insert size has been determined by
Pulsed Field Gel Electrophoresis. Clone size distribution
has been plotted graphically. Clone size distribution for
each segment has been plotted graphically. While
analyzing clones using pulse-field electrophoresis to determine
the average insert size, non-insert clones containing a small
deleted vector fragment consistent with sucrose resistance
were observed. Further in depth characterization of
the library is on going in our lab and data will be updated
on our web page periodically.
Please direct questions
concerning this library to either Pieter
J. de Jong or Kazutoyo
| Ordering &
The library is available
in several formats. Individual clones, and high-density hybridization
filters are obtainable. For ordering and shipping
details, please view the ordering
and pricing information page.
Academic and commercial
users interested in a copy of the BAC library should contact
Pieter J. de Jong ( firstname.lastname@example.org
), fax: (510) 450-7924).
Osoegawa K, Vessere GM, Li Shu C, Hoskins RA, Abad JP, de Pablos B, Villasante A, de Jong PJ. BAC clones generated from sheared DNA. Genomics 2007 89:291-9.